[Lactose intolerance: changing paradigms due to molecular biology].
In most mammals, lactase activity declines on the intestinal wall after weaning, characterizing primary hypolactasia that provokes symptoms of lactose intolerance. The intensity of symptoms of distention, flatulence, abdominal pain and diarrhea varies, according to the amount of ingested lactose, and increases with age. Hypolactasia is genetically determined; nonetheless, a mutation occurred that had made a part of mankind tolerate milk in adulthood. Diagnosis is made by a tolerance test, using the lactose challenge. With the discovery made by the Finns of polymorphism associated with lactase persistence, mainly, in Northern Europe, the genetic test was incorporated as a more comfortable diagnostic tool for the intolerant. In Brazil, 43% of Caucasian and Mulatto groups have lactase persistence allele, with hipolactasia more frequently found among Blacks and Japanese. However, in clinical practice people with hypolactasia may be advised to consume certain dairy products and food containing lactose without developing intolerance symptoms, whereas others will need a lactose restriction diet.


The 10-year cost-effectiveness of lifestyle intervention or metformin for diabetes prevention: an intent-to-treat analysis of the DPP/DPPOS.
The Diabetes Prevention Program (DPP) and its Outcomes Study (DPPOS) demonstrated that either intensive lifestyle intervention or metformin could prevent type 2 diabetes in high-risk adults for at least 10 years after randomization. We report the 10-year within-trial cost-effectiveness of the interventions. Data on resource utilization, cost, and quality of life were collected prospectively. Economic analyses were performed from health system and societal perspectives. Over 10 years, the cumulative, undiscounted per capita direct medical costs of the interventions, as implemented during the DPP, were greater for lifestyle ($4,601) than metformin ($2,300) or placebo ($769). The cumulative direct medical costs of care outside the DPP/DPPOS were least for lifestyle ($24,563 lifestyle vs. $25,616 metformin vs. $27,468 placebo). The cumulative, combined total direct medical costs were greatest for lifestyle and least for metformin ($29,164 lifestyle vs. $27,915 metformin vs. $28,236 placebo). The cumulative quality-adjusted life-years (QALYs) accrued over 10 years were greater for lifestyle (6.81) than metformin (6.69) or placebo (6.67). When costs and outcomes were discounted at 3%, lifestyle cost $10,037 per QALY, and metformin had slightly lower costs and nearly the same QALYs as placebo. Over 10 years, from a payer perspective, lifestyle was cost-effective and metformin was marginally cost-saving compared with placebo. Investment in lifestyle and metformin interventions for diabetes prevention in high-risk adults provides good value for the money spent.


LC-MS/MS quantification of salvinorin A from biological fluids.
A facile method for quantifying the concentration of the powerful and widely available hallucinogen salvinorin A (a selective kappa opioid agonist) from non-human primate cerebrospinal fluid (CSF) and human plasma has been developed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in positive electrospray ionization (ESI) mode. With CSF solid phase extraction can be avoided completely by simply diluting each sample to 10 % (v/v) acetonitrile, 1 % (v/v) formic acid and injecting under high aqueous conditions for analyte focusing. Extensive plasma sample preparation was investigated including protein precipitation, SPE column selection, and plasma particulate removal. Human plasma samples were centrifuged at 21,000 × gravity for 4 minutes to obtain clear particulate-free plasma, from which 300 μl was spiked with internal standard and loaded onto a C18 SPE column with a 100 mg mL-1 loading capacity. Guard columns (C18, hand packed 1 mm × 20 mm) were exchanged after backpressure increased above 4600psi, about 250 injections. A shallow acetonitrile/water gradient was used, 29 to 33% CH3CN over 8 minutes to elute salvinorin A. Reduction of chemical noise was achieved using tandem mass spectrometry with multiple reaction monitoring while sensitivity increases were observed using a 50 μL injection volume onto a small bore analytical column (C18, 1 mm ID × 50 mm) thus increasing peak concentration. Limits of quantification were found to be 0.0125 ng mL-1 (CSF) and 0.05 ng mL-1 (plasma) with interday precision and accuracy below 1.7 % and 9.42 % (CSF) and 3.47 % and 12.37 % (plasma) respectively. This method was used to determine the concentration of salvinorin A from an in vivo Rhesus monkey study and a trial of healthy human research participants, using behaviorally active doses.


Salivary Interleukin-6 Levels among Polycystic Ovary Syndrome Patients with and Without Chronic Periodontitis - A Comparative Study.
Periodontitis is associated with various systemic diseases one of which is poly cystic ovarian syndrome (PCOS). PCOS is a genetically complex endocrinopathy of uncertain etiology affecting women of the reproductive age group which results in the most common cause of anovulatory infertility, menstrual dysfunction, and hirsutism. PCOS has a close association with cardiometabolic risk profile, insulin resistance (IR), hyperinsulinemia, central obesity, dyslipidemia, and increasing the prevalence of cardiovascular risk factors. The common pathway is the chronic low-grade inflammation which is constituted by pro-inflammatory cytokine interleukin (IL)-6. The aim of the study was to compare salivary IL-6 levels among polycystic ovary syndrome (PCOS) patients with and without chronic periodontitis. Newly diagnosed PCOS patients were selected for the study, and the periodontal parameters were recorded. Group A consists of 42 patients of PCOS with periodontitis and Group B consists of 42 patients of PCOS without periodontitis. Salivary levels of IL-6 were compared between the two groups and were assessed by enzyme-linked immunosorbent assay kit (bioassay). The mean pocket depth in Group A was 4.23 ± 0.134 and that of Group B was 1.30 ± 0.06. The mean bleeding on probing in Group A was 1.40 ± 0.40 and in Group B it was 0.91 ± 0.18. The mean clinical attachment level in Group A was 4.87 ± 0.124 and that of Group B was 1.30 ± 0.06. The mean difference in clinical parameters was statistically significant between the groups (P ≤ 0.001). IL-6 level in group A is 102.59 ± 18.2 and in Group B it was 51.3 ± 25.3. Salivary IL-6 levels show a double-fold increase in PCOS with periodontitis than in PCOS without periodontitis. This study reflects the importance of periodontal health and the prevention of periodontal disease so as to minimize IR in PCOS patients with periodontitis.


A novel stenting technique in endoscopic dacryocystorhinostomy.
The objective of our study was to evaluate in a prospective noncomparative case series, the clinical efficacy and the results of modified T-tube insertion instead of silicone tube in endoscopic dacryocystorhinostomy (EDCR). Following ophthalmologic examinations, 22 patients (16 female, 6 male) suffering from recurrent chronic epiphora with postsaccal stenosis for whom DCR was indicated, underwent EDCR at Dumlupinar University Hospital by an otorhinolaryngologist and an ophthalmologist between the years 2006 and 2007. Instead of inserting a classic silicone tube passed through canaliculi and knotted inside the nose, a modified T-tube was used, without passing it through punctum, and was placed to stoma with a less traumatic endonasal endoscopic technique. The medial wall of the sac was incised vertically and the T-tube was placed horizontal to the incision. Postoperatively, the patients were evaluated by endoscopic controls in the first, third, sixth, and twelfth months to evaluate T-tube localization and rhinostomy opening site. The success was defined as resolution of epiphora with patent ostium after evaluation by irrigation and nasal endoscopy. Eighteen patients (82%) showed complete resolution of epiphora at 12-months of follow-up. Re-stenosis was observed only in four cases, one in the sixth and the other in the ninth postoperative months. Granulation tissue formation near the rhinostomy site was seen in one patient, which did not cause any functional problem. The success rate was calculated as 82%. We conclude that modified T-tube placement in EDCR seems to be a practical and less traumatic technique. This is also a cheaper method besides being less traumatic and easier to perform than canalicular stenting. It can be considered as a useful alternative to the classic silicon tubes in cases of epiphora resulting from the postsaccal stenosis.


Optimal management of the critically ill: anaesthesia, monitoring, data capture, and point-of-care technological practices in ovine models of critical care.
Animal models of critical illness are vital in biomedical research. They provide possibilities for the investigation of pathophysiological processes that may not otherwise be possible in humans. In order to be clinically applicable, the model should simulate the critical care situation realistically, including anaesthesia, monitoring, sampling, utilising appropriate personnel skill mix, and therapeutic interventions. There are limited data documenting the constitution of ideal technologically advanced large animal critical care practices and all the processes of the animal model. In this paper, we describe the procedure of animal preparation, anaesthesia induction and maintenance, physiologic monitoring, data capture, point-of-care technology, and animal aftercare that has been successfully used to study several novel ovine models of critical illness. The relevant investigations are on respiratory failure due to smoke inhalation, transfusion related acute lung injury, endotoxin-induced proteogenomic alterations, haemorrhagic shock, septic shock, brain death, cerebral microcirculation, and artificial heart studies. We have demonstrated the functionality of monitoring practices during anaesthesia required to provide a platform for undertaking systematic investigations in complex ovine models of critical illness.


Extending the imaging volume for biometric iris recognition.
The use of the human iris as a biometric has recently attracted significant interest in the area of security applications. The need to capture an iris without active user cooperation places demands on the optical system. Unlike a traditional optical design, in which a large imaging volume is traded off for diminished imaging resolution and capacity for collecting light, Wavefront Coded imaging is a computational imaging technology capable of expanding the imaging volume while maintaining an accurate and robust iris identification capability. We apply Wavefront Coded imaging to extend the imaging volume of the iris recognition application.


A prospective cohort study to examine the association between dietary patterns and depressive symptoms in older Chinese people in Hong Kong.
Dietary patterns are culturally specific and there is limited data on the association of dietary patterns with late-life depression in Chinese. This study examined the associations between dietary patterns and baseline and subsequent depressive symptoms in community-dwelling Chinese older people in Hong Kong. Participants aged ≥65 year participating in a cohort study examining the risk factors for osteoporosis completed a validated food frequency questionnaire at baseline between 2001 and 2003. Factor analysis was used to identify three dietary patterns: "vegetables-fruits" pattern, "snacks-drinks-milk products" pattern, and "meat-fish" pattern. Depressive symptoms were measured at baseline and 4-year using the validated Geriatric Depression Scale. Multiple logistic regression was used for cross-sectional analysis (n = 2,902) to assess the associations between dietary patterns and the presence of depressive symptoms, and for longitudinal analysis (n = 2,211) on their associations with 4-year depressive symptoms, with adjustment for socio-demographic and lifestyle factors. The highest quartile of "vegetables-fruits" pattern score was associated with reduced likelihood of depressive symptoms [Adjusted OR = 0.55 (95% CI: 0.36-0.83), ptrend = 0.017] compared to the lowest quartile at baseline. Similar inverse trend was observed for the highest quartile of "snacks-drinks-milk products" pattern score [Adjusted OR = 0.41 (95% CI: 0.26-0.65), ptrend<0.001] compared to the lowest quartile. There was no association of "meat-fish" pattern with the presence of depressive symptoms at baseline. None of the dietary patterns were associated with subsequent depressive symptoms at 4-year. Higher "vegetables-fruits" and "snacks-drinks-milk products" pattern scores were associated with reduced likelihood of baseline depressive symptoms in Chinese older people in Hong Kong. The longitudinal analyses failed to show any causal relationship between dietary patterns and depressive symptoms in this population.


Effect of prior chronic contractile activity on mitochondrial function and apoptotic protein expression in denervated muscle.
Skeletal muscle is highly adaptable in response to increases and decreases in contractile activity. The purpose of this study was to determine whether the preconditioning of skeletal muscle has a protective effect against subsequent denervation-induced apoptotic protein expression. To investigate this, we chronically stimulated the tibialis anterior and extensor digitorum longus muscles for 7 days (10 Hz, 3 h/day) before 7 days of denervation. Denervation reduced total cytochrome-c oxidase activity by 39%, which was likely a consequence of a decrease in subsarcolemmal (SS) mitochondria. This decrease in the SS subfraction was prevented by prior chronic stimulation and, as a result, maintained total mitochondrial content at control levels. The expression of Bax was elevated 2.2-fold by denervation, and prior chronic stimulation did not attenuate this increase. This produced a increase in the Bax-to-Bcl-2 ratio, indicating greater muscle apoptotic susceptibility. Denervation also decreased state 3 respiration in SS and intermyofibrillar mitochondria and elevated state 4 reactive oxygen species production within both mitochondrial subfractions. These changes were not prevented by prior chronic stimulation. Furthermore, the antioxidant protein MnSOD was also reduced by denervation, whereas Beclin-1 was markedly elevated. This suggests that autophagic cell death could also play a significant part in denervation-induced muscle atrophy. Thus, despite prior chronic stimulation, denervation increases the apoptotic susceptibility of skeletal muscle by altering the Bax-to-Bcl-2 ratio, by increasing reactive oxygen species production, and by reducing the expression of MnSOD. Whether a more extensive stimulation paradigm would be more effective in attenuating apoptosis before muscle disuse remains to be determined.


[Effect of mineral waters with different salt composition on changes in parenchymal-stromal ratio of the laryngeal wall glands in rats].
The aim of the present investigation was to study the laryngeal glandular apparatus as influenced by the application of baths with different mineral composition. 100 mature outbred male rats aged 3-4 months with 180-200 g body mass were studied in this investigation (including 60 experimental and 40 control animals). After a 28-days-long balneologic course using mineral waters with different composition, the histological and morphometric study of laryngeal gland was performed in different parts of the organ to assess the relative contents of parenchyma and stroma. As a result of investigation performed it was demonstrated that course application of iodobromine and bituminous baths resulted in an increased parenchymal contribution to the glandular structure with a corresponding reduction in a relative stromal contents. However, course application of strong (concentrated) sulfide baths causes the stromal outgrowth and glandular epithelium reduction.


One-Step Production Using a Microfluidic Device of Highly Biocompatible Size-Controlled Noncationic Exosome-like Nanoparticles for RNA Delivery.
Size-controlled lipid nanoparticle (LNP)-based DNA/RNA delivery is a leading technology for gene therapies. For DNA/RNA delivery, typically, a cationic lipid is used to encapsulate DNA/RNA into LNPs. However, the use of the cationic lipid leads to cytotoxicity. In contrast, noncationic NPs, such as exosomes, are ideal nanocarriers for DNA/RNA delivery. However, the development of a simple one-step method for the production of size-controlled noncationic NPs encapsulating DNA/RNA is still challenging because of the lack of electrostatic interactions between the cationic lipid and negatively charged DNA/RNA. Herein, we report a microfluidic-based one-step method for the production of size-controlled noncationic NPs encapsulating small interfering RNA (siRNA). Our microfluidic device, named iLiNP, enables the efficient encapsulation of siRNA, as well as control over the NP size, by varying the flow conditions. We applied this method to produce size-controlled exosome-like NPs. The siRNA-loaded exosome-like NPs did not show in vitro cytotoxicity at a high siRNA dosage. In addition, we investigated the effect of the size of the exosome-like NPs on the target gene silencing and found that the 40-50 nm-sized NPs suppressed target protein expression at a dose of 20 nM siRNA. The iLiNP-based one-step production method for size-controlled noncationic-NP-encapsulated RNA is a promising method for the production of artificial exosomes and functionally modified exosomes for gene and cell therapies.


Bile Salt Micelles and Phospholipid Vesicles Present in Simulated and Human Intestinal Fluids: Structural Analysis by Flow Field-Flow Fractionation/Multiangle Laser Light Scattering.
Knowledge about colloidal assemblies present in human intestinal fluids (HIFs), such as bile salt micelles and phospholipid vesicles, is regarded of importance for a better understanding of the in vivo dissolution and absorption behavior of poorly soluble drugs (Biopharmaceutics Classification System class II/IV drugs) because of their drug-solubilizing ability. The characterization of these potential drug-solubilizing compartments is a prerequisite for further studies of the mechanistic interplays between drug molecules and colloidal structures within HIFs. The aim of the present study was to apply asymmetrical flow field-flow fractionation (AF4) in combination with multiangle laser light scattering in an attempt to reveal coexistence of colloidal particles in both artificial and aspirated HIFs and to determine their sizes. Asymmetrical flow field-flow fractionation/multiangle laser light scattering analysis of the colloidal phase of intestinal fluids allowed for a detailed insight into the whole spectrum of submicron- to micrometer-sized particles. With respect to the simulated intestinal fluids mimicking fasted and fed state (FaSSIF-V1 and FeSSIF-V1, respectively), FaSSIF contained one distinct size fraction of colloidal assemblies, whereas FeSSIF contained 2 fractions of colloidal species with significantly different sizes. These size fractions likely represent (1) mixed taurocholate-phospholipid-micelles, as indicated by a size range up to 70 nm (in diameter) and a strong UV absorption and (2) small phospholipid vesicles of 90-210 nm diameter. In contrast, within the colloidal phase of the fasted state aspirate of a human volunteer, 4 different size fractions were separated from each other in a consistent and reproducible manner. The 2 fractions containing large particles showed mean sizes of approximately 50 and 200 nm, respectively (intensity-weighted mean diameter, Dz), likely representing mixed cholate/phospholipid micelles and phospholipid vesicles, respectively. The sizes of the smaller 2 fractions being below the size range of multiangle laser light scattering analysis (<20 nm) and their strong UV absorption indicates that they represent either pure cholate micelles or small mixed micelles. Within the colloidal fraction of the fed-state human aspirate, similar colloidal assemblies were detected as in the fasted state human aspirates. The observed differences between SIF and HIF indicate that the simulated intestinal fluids (FaSSIF-V1 and FeSSIF-V1) represent rather simplified models of the real human intestinal environment in terms of coexisting colloidal particles. It is hypothesized that the different supramolecular assemblies detected differ in their lipid composition, which may affect their affinity toward drug compounds and thus the drug-solubilizing capabilities.


Identification of maize silicon influx transporters.
Maize (Zea mays L.) shows a high accumulation of silicon (Si), but transporters involved in the uptake and distribution have not been identified. In the present study, we isolated two genes (ZmLsi1 and ZmLsi6), which are homologous to rice influx Si transporter OsLsi1. Heterologous expression in Xenopus laevis oocytes showed that both ZmLsi1 and ZmLsi6 are permeable to silicic acid. ZmLsi1 was mainly expressed in the roots. By contrast, ZmLsi6 was expressed more in the leaf sheaths and blades. Different from OsLsi1, the expression level of both ZmLsi1 and ZmLsi6 was unaffected by Si supply. Immunostaining showed that ZmLsi1 was localized on the plasma membrane of the distal side of root epidermal and hypodermal cells in the seminal and crown roots, and also in cortex cells in lateral roots. In the shoots, ZmLsi6 was found in the xylem parenchyma cells that are adjacent to the vessels in both leaf sheaths and leaf blades. ZmLsi6 in the leaf sheaths and blades also exhibited polar localization on the side facing towards the vessel. Taken together, it can be concluded that ZmLsi1 is an influx transporter of Si, which is responsible for the transport of Si from the external solution to the root cells and that ZmLsi6 mainly functions as a Si transporter for xylem unloading.


Incidence, Common Pathogens, and Risk Factors for Infection after Primary Anterior Cruciate Ligament Reconstruction: A Systematic Review.
We sought to assess the current literature to present a comprehensive summary of the incidence, common pathogens, and risk factors for infection after anterior cruciate ligament (ACL) reconstruction. PubMed, CINAHL, EMBASE, and Scopus databases were searched for relevant studies reporting on infection after ACL reconstruction. Two reviewers independently screened the extracted studies for adherence to inclusion and exclusion criteria. Studies were selected if they reported on the incidence of infection, pathogens cultured from infected knees, or risk factors for infection after primary ACL reconstruction. Exclusion criteria consisted of studies with fewer than 100 patients or studies that included revision ACL reconstruction. Fifty studies met the inclusion and exclusion criteria, reporting on a total of 316,214 ACL reconstructions. Included studies evaluated between 123 and 104,255 patients. The overall incidence of infection was 0.60% (0.15-2.44%). The most common pathogens were Staphylococcus aureus, S. epidermidis, and coagulase-negative Staphylococci. Five studies reported that the use of hamstring autograft was a statistically significant risk factor for infection after ACL reconstruction, thus making hamstring autograft the most commonly reported risk factor. Other reported risk factors included male sex, use of immuno-suppressive medications or intraarticular steroid injections, prior knee surgery, and diabetes. Systematic review of the literature revealed that infection after ACL reconstruction remains an infrequent event with an incidence of 0.60% (0.15-2.44%). Furthermore, the most common pathogens are from the Staphylococcus genus of bacteria, comprising 84% of all culture-positive infections. Multiple risk factors have been reported for ACL reconstruction; however, statistical significance varied across studies. Together, these findings may help guide physicians in the prevention and treatment of infection after ACL reconstruction.


Patient- and physician-reported outcomes from two phase 3 randomized studies (RAJ3 and RAJ4) of peficitinib (ASP015K) in Asian patients with rheumatoid arthritis.
Peficitinib (ASP015K), a novel oral Janus kinase inhibitor, has demonstrated efficacy and safety in the treatment of patients with rheumatoid arthritis (RA). This study evaluated the effect of peficitinib on patient- and physician-reported outcomes in Asian patients with RA and an inadequate response to prior disease-modifying antirheumatic drugs (DMARDs). Patients from two randomized, placebo-controlled, double-blind, phase 3 trials (RAJ3 and RAJ4) received once-daily peficitinib 100 mg, peficitinib 150 mg, or placebo, alone or in combination with DMARDs (RAJ3), or in combination with methotrexate (RAJ4). Mean changes in Work Productivity and Activity Impairment (WPAI) questionnaire domain scores from baseline, and percentages of patients achieving minimal clinically important differences (MCIDs) for patient- and physician-reported outcomes (WPAI, Health Assessment Questionnaire - Disability Index [HAQ-DI], and Subject's Global Assessment of Pain [SGAP]), and Physician's Global Assessment of disease activity (PGA) were evaluated at weeks 4, 8, 12, and 12/early termination (ET). Data from 1025 patients were analyzed. At week 12/ET in both studies, patients who received peficitinib 100 mg or 150 mg reported significantly improved WPAI domain scores from baseline (except for absenteeism in RAJ4) compared with placebo (both doses, p<0.05). A higher proportion of peficitinib- versus placebo-treated patients achieved MCID in WPAI, HAQ-DI, SGAP, and PGA in studies RAJ3 and RAJ4. Significant differences with peficitinib versus placebo were evident in both studies as early as week 4 in HAQ-DI (peficitinib 150 mg only), SGAP, and PGA, and week 8 in WPAI loss of work productivity and daily activity impairment. At week 12/ET, significantly higher proportions of patients receiving peficitinib versus placebo achieved MCID in HAQ-DI, SGAP, PGA, and WPAI domains of presenteeism (RAJ3 only), loss of work productivity (RAJ3 only), and daily activity impairment (p<0.05 for all comparisons). Peficitinib 100 mg or 150 mg administered daily over 12 weeks resulted in clinically meaningful improvements in outcomes that are important to RA patients, including pain, physical function, and work productivity and activity. These observations were reinforced through similar improvements in physicians' rating of disease activity. RAJ3: ClinicalTrials.gov, NCT02308163 , registered 4 December 2014. RAJ4: ClinicalTrials.gov, NCT02305849 , registered 3 December 2014.


[Feline sarcoid in a 1-year-old domestic short-haired cat caused by bovine papillomavirus type 14 in Switzerland].
A 1-year- old domestic short haired cat, living on a farm in Switzerland, was presented to the veterinarian with a 5 cm in diameter mass, bulging from her left nostril. The mass was only incompletely removed because of its unfavourable location. Histologically, the lesion consisted of an infiltrative growing spindeloid proliferation in close approximation to the epidermis and was diagnosed as a feline sarcoid tumour. The presence of Bovine Papillomavirus type 14 (BPV-14) specific DNA could be identified in the tissue by using two PCR assays. The amplified sequences of 194 and 549 base pairs (bp) were 99% and 100% identical with a virus isolated after autopsy, from a cat with feline sarcoid in the USA. The cat recovered completely after an even incomplete surgical excision and no recurrence could be observed 10 months later.


Targeting P-glycoprotein for effective oral anti-cancer chemotherapeutics.
Oral anticancer drug treatment represents a significant change to current oncology practice. Support for oral anticancer treatment is driven by issues of pharmacoeconomics, accommodating the need for protracted drug administration for many emerging cytostatic therapies, response to patient preference and in improving patient quality of life. Much focus has concentrated on defining the cellular mechanisms underlying the pharmacokinetic limitations associated with the oral route of administration. However, the potential effects of oral anticancer drugs on gut associated host mediated immunity have been overlooked. Given that the immune system is central for tumour rejection, an assessment of the potential effects oral anticancer drugs may have at this level, and the impact of this on the treatment of gastrointestinal malignancy is of significant clinical importance. P-glycoprotein is a multidrug transporter that contributes to the reduced bioavailability of many orally administered medications. P-glycoprotein achieves this by virtue of its drug efflux capacity at the level of the gut epithelia. P-glycoprotein is also notorious for contributing to the multidrug resistance phenotype observed in many drug refractory human cancers. Likewise, this drug transporter serves a role in the cells of the immune system; particularly in dendritic cell maturation and function. This multifaceted involvement in drug disposition, cancer drug resistance and regulation of the immune response makes P-glycoprotein an attractive target for the optimization of oral anticancer drug treatment strategies. This review introduces and discusses for the first time the potential impact that oral anticancer drugs may have on P-glycoprotein expression and function and the potential consequences of this on dendritic cell function in relation to human cancer. This review also aims to foster a better understanding of the host mediated immunological mechanisms which may be potentially manipulated in cancer patients undergoing oral chemotherapy.


A novel cylindrical microwell featuring inverted-pyramidal opening for efficient cell spheroid formation without cell loss.
Spheroid cultures have been often used to simulate and understand in situ biological occurrences with potential to be further applied to therapeutic approaches, such as cell transplantation. However, traditional lab-scale techniques hardly reached the needed large scale production of cell spheroids, thus limiting their versatility in many biomedical fields. Microscale technologies have rapidly improved in the last decade, and contributed to the large scale production of cell spheroids with high controllability and reproducibility. Nonetheless, the existing microwell culture platforms are problematic due to unwanted cellular adhesion to the substrate as well as due to substantial amounts of cell loss. In this study, we have developed a novel configuration of cylindrical type polyethylene glycol (PEG) hydrogel microwells featuring inverted-pyramidal openings (iPO). Highly refined microstructures of our novel microwell could be fabricated by our optimized micro-electro mechanical system protocols consisting of a silicon (Si) wet/dry etching, Si-to-polydimethylsiloxane substrate bonding, and the established soft-lithography techniques. The iPO, the PEG hydrogel, and the cylindrical geometry of our microwell successfully (1) avoided inefficient washing steps after cell seeding, (2) achieved the complete resistance to cellular adhesion on the microwell substrate, and (3) made all seeded cells readily gathered and jam-packed to form cell spheroids with uniform size, respectively. The maximal sizes of cell spheroids were confined to below 200 μm according to the size of microwells used in this study. The efficiency testing for cell spheroid formation was conducted in comparison with other types of microwells that have been often used in the fields. The results showed that our novel microwell platform effectively reached almost zero percent of cell loss while mass-producing human mesenchymal stem cell spheroids with highly precise control over spheroid's size and cell number. We believe that this study could deliver an effective method to extend the practical usability of cell spheroids in a variety of biomedical applications.


Association between GRK4 and DRD1 gene polymorphisms and hypertension: a meta-analysis.
The role of GRK4 and DRD1 genes in hypertension remains controversial. We performed a meta-analysis to determine whether GRK4 and DRD1 polymorphisms influence the risk of hypertension and examined the relationship between the genetic variances and the etiology of hypertension. Relevant case-control studies were retrieved by database searches and selected according to established inclusion criteria. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated to evaluate the strength of the associations. Meta-regression, subgroup analysis, and sensitivity analysis were performed. A total of 15 articles containing 29 studies were finally included. In the dominant model, rs4532 locus of DRD1 gene was related to hypertension with a pooled OR of 1.353 (95% CI =1.016-1.802, P=0.038). Subgroup analysis for ethnicity showed that rs1024323 locus of GRK4 gene was associated with hypertension in Caucasians (OR =1.826, 95% CI =1.215-2.745, P=0.004) but not in East Asians and Africans. Rs4532 locus was associated with hypertension in East Asians (OR =1.833, 95% CI =1.415-2.376, P,0.001) but not in Caucasians. These data provide possible references for future case-control studies in hypertension.


[Endometrial ablation by liquid heat using the EnAbl system in patients after abdominal hysterectomy. Study of safety].
Security and efficacy of an EnABL, were evaluated; this system had been designed for endometrial ablation by liquid heat in uterine cavity. Eleven patients were included, from the Outpatient Department (INP), programmed for different causes that objectives of this study for abdominal hysterectomy due to abnormal uterine bleeding. The study was approved by Ethical and Scientific Committees. Each patient had preoperative studies, endometrial biopsy, PAP, and ultrasound, in series. The patients with uterus larger than 14 cm, possible cancer; younger than 18 year, or with active bleeding at the time of hysterectomy, were excluded. Each patient received a schema of standard endometrial suppression. The system was applied previously to TAH; 2271 measurements of temperature at one minute intervals were done during liquid heat application. Surgical pieces were sent to histology to analyze the thermic damage through macroscopic aspect, HE tinction and an immunohistochemical cellular viability of NADH test. The thermal damage by macroscopic appearance was 4.33 +/- 1.03 mm, with HE of 4.15 +/- 0.75 mm and with the tinction of NADH of 4.25 mm +/- 0.79. The maximal damage by macroscopic appearance was 4.33 +/- 1.03 mm, with HE of 4.15 +/- 0.75 mm, and with NADH it was 4.25 mm +/- 0.79. The maximal damage was by macroscopic appearance was 6.0 mm and the minimal one was 2.0 mm. In evaluation by H/E, maximal was 5.1 mm and the minimal one was 2.3 mm. In NADH tinction maximal was 4.25 and minimal 2.4 mm. Horizontal analysis showed eight patients with major thermal damage at 4 mm; and two patients with lesser damage at 4 mm. Sub-serous temperatures measured with thermopairs, were done 1504 times, in total. Average was 36.28 degrees C, ranging 35 to 37 degrees C. Maximal temperature was 45 degrees C, 1-2 mm, bellow from serous surface of uterus, and the last one was 28 degrees C. The serous temperatures measured by infrared radiation were done 767 times with an average temperature of 34.6 degrees C. Average was 34-35 C, with standard deviation of 1-2 degrees C. Maximal temperature was 40 degrees C and minimal 29 degrees C. There were no adverse effects. This study shows that EnAbl system is an efficacious method.


VNTR typing studies of Mycobacterium leprae in China: assessment of methods and stability of markers during treatment.
To evaluate the reliability and feasibility of two methods of multilocus variable number of tandem repeat analysis (MLVA) for strain typing of M. leprae, and to study whether short tandem repeat loci are stable and suitable for epidemiological study of leprosy. Total DNA was extracted from skin biopsies of 20 new multibacillary (MB) patients from China diagnosed in 2006. To determine the copy numbers of short tandem repeats (STRs) for 13 loci, we amplified each locus individually by PCR, followed by sequence analysis of the amplicons. Separately, the same loci, plus four others were amplified by Multiplex PCRs (MP) using fluorescent primers and the copy number was identified by fragment length analysis (MP-FLA). MLVA was also performed at different times during treatment for a subset of the patients. Genetic variability of M. leprae in China can be assessed in microsatellite loci. (GTA)9 and (TTC)21 loci are hypervariable, with array sizes of 25 repeat units or more. The expansion of the (GTA)9 locus is a characteristic of some M. leprae isolates in China. A high level of allele concordance was observed between PCR-sequencing and MP-FLA methods. However, MP-FLA method was cost-effective, rapid, high throughput and suitable for strain typing. Five of the 20 isolates of M. leprae were from patients residing in the same township in Qiubei County, Yunnan, and matched closely by MLVA. Three of these patients are family contacts of previously diagnosed patients, with intra-familial strain types being similar, suggesting infections from common sources and transmission chain(s). The VNTR patterns were highly similar in biopsy and slit skin smears (SSS) before treatment, and in the SSS collected at various time points during treatment. Taken together, VNTR strain typing is a useful tool for study of short range transmission in leprosy.


Club cells surviving influenza A virus infection induce temporary nonspecific antiviral immunity.
A brief window of antigen-nonspecific protection has been observed after influenza A virus (IAV) infection. Although this temporary immunity has been assumed to be the result of residual nonspecific inflammation, this period of induced immunity has not been fully studied. Because IAV has long been characterized as a cytopathic virus (based on its ability to rapidly lyse most cell types in culture), it has been a forgone conclusion that directly infected cells could not be contributing to this effect. Using a Cre recombinase-expressing IAV, we have previously shown that club cells can survive direct viral infection. We show here not only that these cells can eliminate all traces of the virus and survive but also that they acquire a heightened antiviral response phenotype after surviving. Moreover, we experimentally demonstrate temporary nonspecific viral immunity after IAV infection and show that surviving cells are required for this phenotype. This work characterizes a virally induced modulation of the innate immune response that may represent a new mechanism to prevent viral diseases.


Degree of N-Methylation of Nucleosides and Metabolites Controls Binding Affinity to Pristine Silica Surfaces.
Biological molecules interact with silica (SiO2) surfaces with binding affinities that greatly vary depending on their physical-chemical properties. However, the quantitative characterization of biological compounds adsorbed on silica surfaces, especially of compounds involved in fast, reversible interactions, has been challenging, and the driving forces are not well understood. Here, we show how carbon-13 NMR spin relaxation provides quantitative atomic-detail information about the transient molecular binding to pristine silica surfaces, represented by colloidally dispersed silica nanoparticles (SNPs). Based on the quantitative analysis of almost two dozen biological molecules, we find that the addition of N-methyl motifs systematically increases molecular binding affinities to silica in a nearly quantitatively predictable manner. Among the studied compounds are methylated nucleosides, which are common in epigenetic signaling in nucleic acids. The quantitative understanding of N-methylation may open up new ways to detect and separate methylated nucleic acids or even regulate their cellular functions.


Optical nonlinearity enhancement with graphene-decorated silicon waveguides.
Broadband on-chip optical frequency combs (OFCs) are important for expanding the functionality of photonic integrated circuits. Here, we demonstrate a huge local optical nonlinearity enhancement using graphene. A waveguide is decorated with graphene by precisely manipulating graphene's area and position. Our approach simultaneously achieves both an extremely efficient supercontinuum and ultra-short pulse generation. With our graphene-decorated silicon waveguide (G-SWG), we have achieved enhanced spectral broadening of femtosecond pump pulses, along with an eightfold increase in the output optical intensity at a wavelength approximately 200 nm shorter than that of the pump pulses. We also found that this huge nonlinearity works as a compressor that effectively compresses pulse width from 80 to 15.7 fs. Our results clearly show the potential for our G-SWG to greatly boost the speed and capacity of future communications with lower power consumption, and our method will further decrease the required pump laser power because it can be applied to decorate various kinds of waveguides with various two-dimensional materials.


[The diagnostic significance of particular immune-dominating proteins of isogenic variants of Bacillus Anthracis.]
The polyclonal mono-specific sera to proteins of toxin and antigens of S-layer of Bacillus Anthracis separated in preparative electrophoresis. While fluorescent antibody technique was applied, immunoglobulins of sera to protein of S-layer of Bacillus Anthracis m.m. 94 kDa had specificity and specific activity in presented number of strains. In the soil samples spores in concentration of 1x104 KOE/ml were detected that permits to recommend the given immunoglobulins for indexation of Bacillus Anthracis. The proteins m.m. 90 kDa relevant to antigens of toxin of Bacillus Anthracis reacted in immuno-blotting with sera from patients with skin form of anthrax and guinea pigs immunized and survived after infection. Thew sera to them are species specified and can be applied for diagnostic of disease (detection of protective antigen) and detection of production of proteins in reaction of immunodiffusion with growing cultures.


Digestion of soybean meal and canola meal protein and amino acids in the digestive tract of young ruminants.
Eight male Holstein calves (body weight 68 +/- 5 kg; age 75 +/- 6 d), each with a permanent re-entrant pancreatic cannula and T-type ileal and duodenal cannulas, were used in a crossover design with four animals per group to determine amino acid kinetics and digestibilities in the digestive tract of calves fed soybean meal (SBM) and canola meal (CM) protein. The SBM and CM diets were fed twice daily at a level of 900 g at each feeding time (air-dry basis). With the exception of methionine, crude protein and amino acid flows at the proximal duodenum, expressed as a percentage of intake, were not influenced by dietary protein source. Apparent ileal and total tract digestibilities of CP and amino acids were reduced (P less than .05) by feeding CM compared to SBM, but apparent ileal digestibility of methionine was not affected by dietary protein source. Except for methionine, net disappearance of all amino acids in the small intestine, relative to the amount fed, was higher for the SBM diet than for the CM diet. Net disappearance or synthesis of amino acids in the large intestine were not affected by dietary protein source. Similarly, dietary protein source did not affect (P greater than .05) the secretion of pancreatic juice or concentrations of protein, chymotrypsin and trypsin in pancreatic juice. Soybean meal protein has higher ileal and total gastrointestinal tract digestibility than CM protein for young, growing calves.


Management of HIV infection in adults.
The natural history of HIV infection continues to change with improved diagnostic and therapeutic modalities available to manage opportunistic infections and malignancies. Antiretroviral therapy with zidovudine and other investigational agents has improved the median survival of AIDS patients from 11 months in 1985 to 18-25 months at present. Most importantly, early intervention with zidovudine can delay onset of clinical illness in asymptomatic patients and progression to AIDS in symptomatic patients. A 500 mg/d dose has been found as effective as previously recommended doses of 1200-1500 mg/day. Lower doses decrease the incidence and severity of adverse effects and therapeutic benefit appears to be greatest in asymptomatic patients with CD4 lymphocyte counts less than 500/ul. Indications for zidovudine, therefore, have been expanded to include asymptomatic adults with CD4 lymphocyte counts less than 500/ul. Concerning early intervention with zidovudine, studies were not designed to measure survival or define the optimal timing of intervention based on immunologic status. In addition, long-term benefits are not clearly defined, particularly since the drug seems to lose clinical effectiveness after approximately two years, probably due to emergence of resistant HIV strains. Adverse effects continue to occur even at low doses including headaches, nausea, anemia and neutropenia, myopathy and possible hepatitis. Nevertheless, the overall clinical benefit seems to be greatest, albeit temporary, in asymptomatic patients. The optimal dosage appears to be 500-600 mg/d; however, this may not be sufficient for infection in the central nervous system.


Genome-scale classification of metabolic reactions and assignment of EC numbers with self-organizing maps.
The automatic perception of chemical similarities between metabolic reactions is required for a variety of applications ranging from the computer-aided validation of classification systems, to genome-scale reconstruction (or comparison) of metabolic pathways, to the classification of enzymatic mechanisms. Comparison of metabolic reactions has been mostly based on Enzyme Commission (EC) numbers, which are extremely useful and widespread, but not always straightforward to apply, and often problematic when an enzyme catalyzes several reactions, when the same reaction is catalyzed by different enzymes, when official full EC numbers are unavailable or when reactions are not catalyzed by enzymes. Different methods should be available to compare metabolic reactions. Simultaneously, methods are required for the automatic assignment of EC numbers to reactions still not officially classified. We have proposed the MOLMAP reaction descriptors to numerically encode the structural transformations resulting from a chemical reaction. Here, such descriptors are applied to the mapping of a genome-scale database of almost 4000 metabolic reactions by Kohonen self-organizing maps (SOMs), and its screening for inconsistencies in EC numbers. This approach allowed for the SOMs to assign EC numbers at the class, subclass and sub-subclass levels for reactions of independent test sets with accuracies up to 92, 80 and 70%, respectively. Different levels of similarity between training and test sets were explored. The approach also led to the identification of a number of similar reactions bearing differences at the EC class level. The programs to generate MOLMAP descriptors from atomic properties included in SDF files are available upon request for evaluation.


A preliminary study of the morphology of the upper gum pad at the age of 6 months.
Longitudinal records of 109 individuals born at full term and 30 pairs of twins have been used to study factors which determine the form of the upper gum pad at the age of six months and before the emergence of teeth. The contribution of heredity was found to be less than at birth, and that derived from the underlying teeth to have increased during the first six months of life. It is concluded that the shape of the alveolar process may be modified by physical forces such as sucking habits before teeth erupt into the mouth.


Divergent engagements between adeno-associated viruses with their cellular receptor AAVR.
Adeno-associated virus (AAV) receptor (AAVR) is an essential receptor for the entry of multiple AAV serotypes with divergent rules; however, the mechanism remains unclear. Here, we determine the structures of the AAV1-AAVR and AAV5-AAVR complexes, revealing the molecular details by which PKD1 recognizes AAV5 and PKD2 is solely engaged with AAV1. PKD2 lies on the plateau region of the AAV1 capsid. However, the AAV5-AAVR interface is strikingly different, in which PKD1 is bound at the opposite side of the spike of the AAV5 capsid than the PKD2-interacting region of AAV1. Residues in strands F/G and the CD loop of PKD1 interact directly with AAV5, whereas residues in strands B/C/E and the BC loop of PKD2 make contact with AAV1. These findings further the understanding of the distinct mechanisms by which AAVR recognizes various AAV serotypes and provide an example of a single receptor engaging multiple viral serotypes with divergent rules.


Functional recovery of denervated skeletal muscle with sensory or mixed nerve protection: a pilot study.
Functional recovery is usually poor following peripheral nerve injury when reinnervation is delayed. Early innervation by sensory nerve has been indicated to prevent atrophy of the denervated muscle. It is hypothesized that early protection with sensory axons is adequate to improve functional recovery of skeletal muscle following prolonged denervation of mixed nerve injury. In this study, four groups of rats received surgical denervation of the tibial nerve. The proximal and distal stumps of the tibial nerve were ligated in all animals except for those in the immediate repair group. The experimental groups underwent denervation with nerve protection of peroneal nerve (mixed protection) or sural nerve (sensory protection). The experimental and unprotected groups had a stage II surgery in which the trimmed proximal and distal tibial nerve stumps were sutured together. After 3 months of recovery, electrophysiological, histological and morphometric parameters were assessed. It was detected that the significant muscle atrophy and a good preserved structure of the muscle were observed in the unprotected and protective experimental groups, respectively. Significantly fewer numbers of regenerated myelinated axons were observed in the sensory-protected group. Enhanced recovery in the mixed protection group was indicated by the results of the muscle contraction force tests, regenerated myelinated fiber, and the results of the histological analysis. Our results suggest that early axons protection by mixed nerve may complement sensory axons which are required for promoting functional recovery of the denervated muscle natively innervated by mixed nerve.


Reduction of iodinated contrast load with the renal artery catheterization technique during endovascular aortic repair.
We aimed to present our clinical experience with the renal artery catheterization (RAC) technique, which reduces the volume of intra-arterial contrast media (ICM) used during endovascular aortic repair (EVAR), and describe the short-term results of this technique. We retrospectively evaluated 16 patients (15 males and one female) who underwent EVAR between March 2011 and February 2012 using the RAC technique for an abdominal aortic aneurysm. A Simmons-1 catheter was preferred for renal artery cannulation. The mean age of the patients at the time of treatment was 70 years (range, 61-82 years). Fifteen cases were fusiform aneurysms, and one case was a saccular aneurysm. Creatinine and estimated glomerular filtration rate (eGFR) values were recorded before the procedure and during the first 72 hours postprocedure. Bifurcated stent grafts were implanted with 100% procedural success using the RAC technique. The inferiorly positioned renal artery was cannulated with a Simmons-1 catheter in the first five patients, and was maintained at the level of the renal artery orifice in the remaining patients. The mean volume of the ICM used was 47 mL (range, 23-83 mL). The creatinine and eGFR values were not significantly different between the pre- and postoperative periods (P > 0.05). Reducing the volume of ICM used during EVAR is critical for protecting renal function. The RAC technique is a safe and effective method in appropriate patients when performed by experienced clinicians.


Effect of rosuvastatin on dyslipidemia and other parameters associated with metabolic syndrome in Saudi patients.
Metabolic syndrome (MS) is a constellation of metabolic irregularities consisting of dyslipidemia, hypertension, hyperglycemia, chronic inflammatory, and hypercoagulable state predisposing to diabetes and cardiovascular events. Statins are first-line drugs to treat the associated atherogenic dyslipidemia. Effect of rosuvastatin on MS in Saudi patients was studied. Prospective, open label, randomized clinical study. Patients of either sex ≥18 years (n = 153) having MS as per modified National Cholesterol Education Program Adult Treatment Panel III criteria were prescribed rosuvastatin 10 mg OD for 24 weeks. Serum lipids, biochemical, clinical, and anthropometric parameters were studied before and after treatment. Statistical Package for Social Sciences version17 was used. Descriptive analysis was used for all variables and documented as mean ± SD. Normality checked by Shapiro-Wilk test, Kurtosis and Skewness Z-score, and visualization of histograms. Lipid levels and other parameters before and after treatment were evaluated by paired t-test for parametric data and Wilcoxon signed rank test for nonparametric data. Pre- and post-test values were correlated by Pearson's correlation coefficient. Multiple regression analysis was performed to see effect of other variables. Highly significant reduction was observed in low density lipoprotein cholesterol, total cholesterol, triglycerides; very low density lipoprotein cholesterol, non-high density lipoprotein cholesterol and atherosclerotic index with an elevation in high density lipoprotein cholesterol. A total of 86% patients reached low density lipoprotein cholesterol goal of ≤ 100 mg/dL. Beneficial response was observed on other associated parameters. There was strong correlation between pre- and post values. No significant effect was observed for any of the variables on cholesterol reduction. No serious/severe adverse effect was observed. Rosuvastatin markedly improved atherogenic dyslipidemia of MS.


Analysis of Competing HIV-1 Splice Donor Sites Uncovers a Tight Cluster of Splicing Regulatory Elements within Exon 2/2b.
The HIV-1 accessory protein Vif is essential for viral replication by counteracting the host restriction factor APOBEC3G (A3G), and balanced levels of both proteins are required for efficient viral replication. Noncoding exons 2/2b contain the Vif start codon between their alternatively used splice donors 2 and 2b (D2 and D2b). For vif mRNA, intron 1 must be removed while intron 2 must be retained. Thus, splice acceptor 1 (A1) must be activated by U1 snRNP binding to either D2 or D2b, while splicing at D2 or D2b must be prevented. Here, we unravel the complex interactions between previously known and novel components of the splicing regulatory network regulating HIV-1 exon 2/2b inclusion in viral mRNAs. In particular, using RNA pulldown experiments and mass spectrometry analysis, we found members of the heterogeneous nuclear ribonucleoparticle (hnRNP) A/B family binding to a novel splicing regulatory element (SRE), the exonic splicing silencer ESS2b, and the splicing regulatory proteins Tra2/SRSF10 binding to the nearby exonic splicing enhancer ESE2b. Using a minigene reporter, we performed bioinformatics HEXplorer-guided mutational analysis to narrow down SRE motifs affecting splice site selection between D2 and D2b. Eventually, the impacts of these SREs on the viral splicing pattern and protein expression were exhaustively analyzed in viral particle production and replication experiments. Masking of these protein binding sites by use of locked nucleic acids (LNAs) impaired Vif expression and viral replication.IMPORTANCE Based on our results, we propose a model in which a dense network of SREs regulates vif mRNA and protein expression, crucial to maintain viral replication within host cells with varying A3G levels and at different stages of infection. This regulation is maintained by several serine/arginine-rich splicing factors (SRSF) and hnRNPs binding to those elements. Targeting this cluster of SREs with LNAs may lead to the development of novel effective therapeutic strategies.


Arginine auxotrophic gene signature in paediatric sarcomas and brain tumours provides a viable target for arginine depletion therapies.
Paediatric sarcomas and brain tumours, remain cancers of significant unmet need, with a poor prognosis for patients with high risk disease or those who relapse, and significant morbidities from treatment for those that survive using standard treatment approaches. Novel treatment strategies, based on the underlying tumour biology, are needed to improve outcomes. Arginine is a semi-essential amino acid that is imported from the extracellular microenvironment or recycled from intracellular precursors through the combined expression of the enzymes ornithine transcarbamylase (OTC), argininosuccinate synthase (ASS) and argininosuccinate lyase (ASL) enzymes. The failure to express at least one of these recycling enzymes makes cells reliant on extracellular arginine - a state known as arginine auxotrophism. Here we show in large in silico patient cohorts that paediatric sarcomas and brain tumours express predominately the arginine transporter SLC7A1 and the arginine metabolising enzyme Arginase 2 (ARG2), but have low-absent expression of OTC. The arginine metabolic pathway correlated with the expression of genes associated with tumour pathogenesis, and overall survival in paediatric sarcomas. This gene signature of arginine auxotrophism indicates paediatric sarcomas and brain tumours are a viable target for therapeutic arginase drugs under current clinical trial development.


Linnorm: improved statistical analysis for single cell RNA-seq expression data.
Linnorm is a novel normalization and transformation method for the analysis of single cell RNA sequencing (scRNA-seq) data. Linnorm is developed to remove technical noises and simultaneously preserve biological variations in scRNA-seq data, such that existing statistical methods can be improved. Using real scRNA-seq data, we compared Linnorm with existing normalization methods, including NODES, SAMstrt, SCnorm, scran, DESeq and TMM. Linnorm shows advantages in speed, technical noise removal and preservation of cell heterogeneity, which can improve existing methods in the discovery of novel subtypes, pseudo-temporal ordering of cells, clustering analysis, etc. Linnorm also performs better than existing DEG analysis methods, including BASiCS, NODES, SAMstrt, Seurat and DESeq2, in false positive rate control and accuracy.


Renoprotection with pituitary adenylate cyclase-activating polypeptide in cyclosporine A-induced nephrotoxicity.
Acute and long-term nephrotoxicity is the major dose-limiting factor for cyclosporine A (CsA). We evaluated the protective effects of pituitary adenylate cyclase-activating polypeptide (PACAP)38 on CsA-induced nephrotoxicity in human renal proximal tubule epithelial (human kidney-2) cells and in intact mice. Confluent (human kidney-2 cells were exposed to CsA (25-50 μmol/L) in the presence or absence of PACAP38 or vasoactive intestinal peptide (10(-10) to 10(-6) M). For studies in vivo, male BALB/c mice (n = 5 in each group) were given a single intraperitoneal injection of CsA (5 mg/kg body weight). Treatment group received 20 μg of PACAP38 2 hours before exposure to CsA and additional doses daily for 10 days. Cyclosporine A caused oxidative injury, marked morphological alterations, apoptosis, and increased expression of transforming growth factor (TGF)-β1 in cell cultures. Pituitary adenylate cyclase-activating polypeptide 38 at 10(-8) mol/L restored cell confluency, reduced TGF-β1 secretion, and preserved cell integrity. In mice, CsA caused tubular injury characterized by loss of tubular epithelial cell brush border membranes, tubular collapse, cellular necrosis, interstitial fibrosis, increased production of TGF-β1, and elevated serum creatinine (3.39 ± 0.21 vs 0.13 ± 0.02 mg/dL in controls, P < 0.01). Treatment with PACAP38 reduced TGF-β1 and tumor necrosis factor-α production in kidney, prevented epithelial-mesenchymal transition of the renal cells, and reduced serum creatinine levels to 1.01 ± 0.18 mg/dL, P < 0.01 versus CsA group. Pituitary adenylate cyclase-activating polypeptide 38 ameliorated renal tubular injury, reduced oxidative injury, and inhibited the expression of TGF-β1 in CsA-exposed murine kidneys. Pituitary adenylate cyclase-activating polypeptide could be a novel renoprotective and antifibrotic agent for CsA nephrotoxicity.


Effects of dietary fish oil supplementation on membrane fluidity and enzyme activity in rat small intestine.
Rats were fed either a fat-free diet supplemented with 10% menhaden oil or a control diet for four months. Intestinal brush border membranes were isolated; phospholipid fatty acid analysis revealed that the membranes from the fish-oil fed animals had higher levels of palmitoleic (C16:1) and eicosapentaenoic (C20:5) acids and lesser levels of stearic (C18:0) linoleic (C18:2) acids compared with controls. The membranes from the fish-oil fed animals had increased levels of alkaline phosphatase activity compared with controls but disaccharidase levels were equivalent in the two groups. Rocket immunoelectrophoresis studies revealed that the increase in alkaline phosphatase activity was due to an increase in the specific activity of the enzyme rather than an increase in the amount of enzyme. Membrane fluidity was assessed by fluorescence anisotropy using diphenylhexatriene and 12-anthroyl stearate as fluorescent probes. The anisotropy of both probes was similar in the two membranes. These studies indicate that fish-oil supplementation alters the fatty acid composition of the intestinal brush border membrane and increases alkaline phosphatase activity without affecting membrane fluidity. Thus the effects of changes in membrane lipid composition on alkaline phosphatase activity appear to result from changes in the local lipid environment of the enzyme rather than from changes in the biophysical characteristics of the membrane.


Outcome of papillary versus clear cell renal cell carcinoma varies significantly in non-metastatic disease.
Renal cell carcinoma (RCC) comprises a heterogenous group of tumors. Traditionally, papillary RCC (pRCC) is associated with a favorable outcome compared to clear cell RCC (ccRCC), while other series report equivalent or worse prognosis. In this paper we comparatively evaluate outcome of pRCC versus ccRCC in two large multi-institutional databases (cohort study), including distribution of pRCC subtypes 1 and 2. Retrospective data of 1,943 surgically treated pRCC patients from 17 European/ North American centers between 1984-2015 were compared to 5,600 ccRCC patients from a database comprising 11 European/ North American centers (1984-2011). Median follow-up was 64.6 months. Differences between pRCC, subtypes, and ccRCC were compared with t-tests, Chi^2-tests, and exact Fisher tests. Cancer-specific mortality was analyzed with cumulative incidence curves and Cox cause-specific hazard models. The robustness of our results was examined with sensitivity analyses. We present that cancer-specific mortality rates and variables as stage, lymph node, and distant metastasis differ significantly between groups. Furthermore, we demonstrate that patients with non-metastatic pRCC had a significantly better cancer-specific mortality (HR 0.76, p = 0.007), when compared to ccRCC. Additionally, pRCC type 2 versus ccRCC exhibited no difference in cancer-specific mortality (HR 0.9, p = 0.722), whereas pRCC type 1 versus ccRCC displayed a risk of death reduced by 69% (p = 0.044). Taken together, outcome of pRCC versus ccRCC varies significantly in non-metastatic disease. Furthermore, pRCC type 2 exhibited no difference in cancer-specific mortality, whereas pRCC type 1 displayed a significantly reduced risk of death. Consequently, there is urgent need to respect histopathological entities and their subtypes, when assigning follow-up or targeted therapy to RCC patients.


Separation of rat muscle aminopeptidases.
By means of chromatography on DEAE-Sephadex, two arylamidases (hydrolysing L-arginine 2-naphthylamide) and three dipeptidyl peptidases (hydrolysing dipeptide 2-naphthylamides) were distinguished in extracts of rat muscle. However, the arylamidase from the larger peak also hydrolysed the dipeptide 2-naphthylamides. Glycyl-L-arginine amide, an alternative substrate for dipeptidyl peptidase I, was not hydrolysed by arylamidase. L-Leucine amide was hydrolysed by an enzyme, presumed to be leucine aminopeptidase, from a separate peak, but was also hydrolysed by arylamidase. Arylamidase, dipeptidyl peptidase III and most of the leucine aminopeptidase could be extracted from the muscle with a neutral salt solution, but dipeptidyl peptidase I was extracted only in the presence of Triton X-100; dipeptidyl peptidase II showed an intermediate extraction behaviour.


Anti-MDA5 antibody-positive dermatomyositis: pathogenesis and clinical progress.
Anti-melanoma differentiation-associated protein 5 (MDA5) antibody-positive dermatomyositis (MDA5-DM) is a subtype of dermatomyositis. Although the aetiology and pathology remain unclear, increasing evidence suggests that viral infection is a potential trigger of MDA5-DM. Multiple factors, including T cells, B cells, neutrophils and macrophages, are implicated in the pathophysiology of MDA5-DM. Distinctive skin rashes, rapidly progressive interstitial lung disease, peripheral lymphopenia and elevated serum ferritin levels are the most prominent clinical and laboratory features of MDA5-DM. Concomitant infection is a common complication of MDA5-DM. The proper evaluation of patients with MDA5-DM requires knowledge of the disease heterogeneity and clinical course variability. Several biomarkers, including serum levels of anti-MDA5 antibodies and biomarkers related to macrophage activation, have been identified as useful tools for monitoring disease activity and prognosis. MDA5-DM shows a poor response to conventional glucocorticoid and immunosuppressant therapy and has a poor overall prognosis. Therefore, there is an urgent need to explore the key pathogenic mechanisms of MDA5-DM and develop novel therapeutic options for patients. This Review discusses recent clinical progress and pathogenic findings of MDA5-DM.


The use of foetal, calf and adult bovine sera for the growth of serially subcultivated diploid cells.
Medium CMRL-1969 supplemented with either foetal calf, calf or adult bovine sera, or a combination of the various sera, was used to subcultivate diploid cells for the preparation of substrates for vaccine production. Supplementation with calf or adult bovine sera gave satisfactory cell yields for at least 3-4 population doublings. The finite lifespan of the diploid cells grown in media supplemented with foetal calf was always longer than with calf or adult bovine sera. However, a combined supplement of adult serum plus a low concentration of foetal calf serum gave results equally as good as foetal calf serum alone. Sera obtained aseptically from adult animals kept in isolation and monitored constantly for health are more desirable than sera obtained from abattoirs. Selection of donor animals and pooling of certain serum batches were found to be important for the preparation of serum supplements. The final protein concentration of the cell culture medium should not exceed 0.8% when adult bovine sera are used. For the growth of WI-38 or MRC-5 cells the use of foetal calf serum can be reduced or eliminated.


Soil Inorganic Carbon Sequestration Following Afforestation Is Probably Induced by Pedogenic Carbonate Formation in Northwest China.
In arid and semiarid areas, the effects of afforestation on soil organic carbon (SOC) have received considerable attention. In these areas, in fact, soil inorganic carbon (SIC), rather than SOC, is the dominant form of carbon, with a reservoir approximately 2-10 times larger than that of SOC. A subtle fluctuation of SIC pool can strongly alter the regional carbon budget. However, few studies have focused on the variations in SIC, or have used stable soil carbon isotopes to analyze the reason for SIC variations following afforestation in degraded semiarid lands. In the Mu Us Desert, northwest China, we selected a shifting sand land (SL) and three nearby forestlands (Populus alba) with ages of 8 (P-8), 20 (P-20) and 30 (P-30) years, and measured SIC, SOC, soil organic and inorganic δ13C values (δ13C-SOC and δ13C-SIC) and other soil properties. The results showed that SIC stock at 0-100 cm in SL was 34.2 Mg ha-1, and it increased significantly to 42.5, 49.2, and 68.3 Mg ha-1 in P-8, P-20, and P-30 lands, respectively. Both δ13C-SIC and δ13C-SOC within the 0-100 cm soil layer in the three forestlands were more negative than those in SL, and gradually decreased with plantation age. Afforestation elevated soil fine particles only at a depth of 0-40 cm. The entire dataset (260 soil samples) exhibited a negative correlation between δ13C-SIC and SIC content (R2 = 0.71, P < 0.01), whereas it showed positive correlation between SOC content and SIC content (R2 = 0.52, P < 0.01) and between δ13C-SOC and δ13C-SIC (R2 = 0.63, P < 0.01). However, no correlation was observed between SIC content and soil fine particles. The results indicated that afforestation on shifting SL has a high potential to sequester SIC in degraded semiarid regions. The contribution of soil fine particle deposition by canopy to SIC sequestration is limited. The SIC sequestration following afforestation is very probably caused by pedogenic carbonate formation, which is closely related to SOC accumulation. Our findings suggest that SIC plays an important role in the carbon cycle in semiarid areas and that overlooking this carbon pool may substantially lead to underestimating carbon sequestration capacity following vegetation rehabilitation.


Two distinct and independent mitochondrial targeting signals function in the sorting of an inner membrane protein, cytochrome c1.
Proteins of the mitochondrial inner membrane display a wide variety of orientations, many spanning the membrane more than once. Some of these proteins are synthesized with NH2-terminal cleavable targeting sequences (presequences) whereas others are targeted to mitochondria via internal signals. Here we report that two distinct mitochondrial targeting signals can be present in precursors of inner membrane proteins, an NH2-terminal one and a second, internal one. Using cytochrome c1 as a model protein, we demonstrate that these two mitochondrial targeting signals operate independently of each other. The internal targeting signal, consisting of a transmembrane segment and a stretch of positively charged amino acid residues directly following it, initially directs the translocation of the preprotein into the intermembrane space. It then inserts into the inner membrane from the intermembrane space side in a delta psi-dependent manner and thereby determines the orientation the protein attains in the inner membrane. Analysis of a number of other presequence-containing protein of the inner membrane suggest that they too contain such internal targeting signals.


Information Length Analysis of Linear Autonomous Stochastic Processes.
When studying the behaviour of complex dynamical systems, a statistical formulation can provide useful insights. In particular, information geometry is a promising tool for this purpose. In this paper, we investigate the information length for n-dimensional linear autonomous stochastic processes, providing a basic theoretical framework that can be applied to a large set of problems in engineering and physics. A specific application is made to a harmonically bound particle system with the natural oscillation frequency ω, subject to a damping γ and a Gaussian white-noise. We explore how the information length depends on ω and γ, elucidating the role of critical damping γ=2ω in information geometry. Furthermore, in the long time limit, we show that the information length reflects the linear geometry associated with the Gaussian statistics in a linear stochastic process.


QoS-Aware Smart Phone-Based User Tracking Application to Prevent Outbreak of COVID-19 in Saudi Arabia.
Diverse variants of COVID-19 are repeatedly making everyday living unstable. In reality, the conclusive retort of this highly contagious virus still is in incognito mode. The health experts' primary guideline on the possible prevention of this disease outbreak, including a list of restrictions and confinements, is insufficient in case of any public congregation. As a result, the demand for precise and upgraded real-time COVID-19 tracking and prevention-based applications increases. However, most of the existing android-based applications face a lack of data security and reliability that cannot satisfy the additional quality of service (QoS) requirements. This paper proposes an easy-to-operate android-based multifunctional application to track individuals' health situations, allow uploading scanning report by the authorized organization like universities, mosques, school, and hospitals and helps the users to maintain guidelines via manageable steps. This article offers a three-layered QoS aware service-oriented task scheduling model upon multitasking android-based frontend focusing the cognitive-based AI applications in healthcare with a continual learning paradigm. Designed model is competent to optimize heterogeneous service scheduling and can minimize data delivery time, as well as the resource cost.


Serological and molecular characteristics of Vibrio vulnificus biotype 3: evidence for high clonality.
Vibrio vulnificus biotype 3 has been implicated as the causative pathogen of an ongoing disease outbreak that erupted in Israel in 1996. Recent work based on multi-locus sequence typing (MLST) showed that V. vulnificus biotype 3 is genetically homogeneous. The aim of this study was to investigate the existence of subpopulations within this homogeneous biotype by characterizing the surface antigens and analysing the sequence diversity of selected outer-membrane protein (OMP)-encoding genes. Rabbit antisera were prepared against biotype 1, 2 and 3 strains. The results of the slide-agglutination test, dot-blot assay (using fresh and boiled cells), and immunoblotting of lipopolysaccharides (LPS) and OMPs were evaluated. By slide-agglutination and dot-blot assays all biotype 3 strains agglutinated with the selected biotype 3 strain. This homogeneity was supported by immunoblot analysis of the LPS. Analysis of OMP patterns revealed that all three biotypes share a considerable number of common bands that are antigenically related. Cluster analysis of DNA sequence data from selected OMP-encoding genes showed that biotype 3 strains form a genetically distinct and homogeneous clone. The homogeneity of surface antigens and the lack of any sequence diversity among both housekeeping and OMP-encoding genes reaffirms the highly clonal nature of biotype 3 and suggests that it has only recently descended from the parent population of V. vulnificus.


Leukocyte Derived Microvesicles as Disease Progression Biomarkers in Slow Progressing Amyotrophic Lateral Sclerosis Patients.
The lack of biomarkers in Amyotrophic Lateral Sclerosis (ALS) makes it difficult to determine the stage of the disease in patients and, therefore, it delays therapeutic trials. Microvesicles (MVs) are possible biomarkers implicated in physiological and pathological functions, however, their role in ALS remains unclear. We investigated whether plasma derived microvesicles could be overrepresented in a group of 40 patients affected by ALS compared to 28 Alzheimer's Disease (AD) patients and 36 healthy volunteers. Leukocyte derived MVs (LMVs) compared to endothelial, platelet, erythrocyte derived MVs, were mostly present in ALS patients compared to AD patients and healthy donors. Correlation analysis corrected for the presence of confounding variables (riluzole, age at onset, site of onset, gender) was tested between PRL (Progression Rate at the Last visit) and LMVs, and a statistically significant value was found (Pearson partial correlation r = 0.407, p = 0.006). We also investigated SOD1, TDP-43 intravesicular protein level in LMVs. Misfolded SOD1 was selectively transported by LMVs and its protein level was associated with the percentage of LMVs in slow progressing patients (r = 0.545, p = 0.033). Our preliminary findings suggest that LMVs are upregulated in ALS patients and they can be considered possible markers of disease progression.


Genomic signatures of extensive inbreeding in Isle Royale wolves, a population on the threshold of extinction.
The observation that small isolated populations often suffer reduced fitness from inbreeding depression has guided conservation theory and practice for decades. However, investigating the genome-wide dynamics associated with inbreeding depression in natural populations is only now feasible with relatively inexpensive sequencing technology and annotated reference genomes. To characterize the genome-wide effects of intense inbreeding and isolation, we performed whole-genome sequencing and morphological analysis of an iconic inbred population, the gray wolves (Canis lupus) of Isle Royale. Through population genetic simulations and comparison with wolf genomes from a variety of demographic histories, we find evidence that severe inbreeding depression in this population is due to increased homozygosity of strongly deleterious recessive mutations. Our results have particular relevance in light of the recent translocation of wolves from the mainland to Isle Royale, as well as broader implications for management of genetic variation in the fragmented landscape of the modern world.


Molecular cloning and characterization of L-galactose-1-phosphate phosphatase from tobacco (Nicotiana tabacum).
L-Galactose-1-phosphate phosphatase (GPPase) is an enzyme involved in ascorbate biosynthesis in higher plants. We isolated a cDNA encoding GPPase from tobacco, and named it NtGPPase. The putative amino acid sequence of NtGPPase contained inositol monophosphatase motifs and metal binding sites. Recombinant NtGPPase hydrolyzed not only L-galactose-1-phosphate, but also myo-inositol-1-phosphate. The optimum pH for the GPPase activity of NtGPPase was 7.5. Its enzyme activity required Mg2+, and was inhibited by Li+ and Ca2+. Its fluorescence, fused with green fluorescence protein in onion cells and protoplasts of tobacco BY-2 cells, was observed in both the cytosol and nucleus. The expression of NtGPPase mRNA and protein was clearly correlated with L-ascorbic acid (AsA) contents of BY-2 cells during culture. The AsA contents of NtGPPase over expression lines were higher than those of empty lines at 13 d after subculture. This suggests that NtGPPase contributes slightly to AsA biosynthesis.


Genetic counseling in Usher syndrome: linkage and mutational analysis of 10 Colombian families.
Usher Syndrome (US), an autosomal recessive disease, is characterized by retinitis pigmentosa (RP), vestibular dysfunction, and congenital sensorineural deafness. There are three recognized clinical types of the disorder. In order to improve genetic counseling for affected families, we conducted linkage analysis and DNA sequencing in 10 Colombian families with confirmed diagnosis of US (4 type I and 6 type II). Seventy-five percent of the US1 families showed linkage to locus USH1B, while the remaining 25% showed linkage to loci USH1B and USH1C. Among families showing linkage to USH1B we found two different mutations in the MYO7A gene: IVS42-26insTTGAG in exon 43 (heterozygous state) and R634X (CGA-TGA) in exon 16 (homozygous state). All six US2 families showed linkage to locus USH2A. Of them, 4 had c.2299delG mutation (1 homozygote state and 3 heterozygous); in the remaining 2 we did not identify any pathologic DNA variant. USH2A individuals with a 2299delG mutation presented a typical and homogeneous retinal phenotype with bilateral severe hearing loss, except for one individual with a heterozygous 2299delG mutation, whose hearing loss was asymmetric, but more profound than in the other cases. The study of these families adds to the genotype-phenotype characterization of the different types and subtypes of US and facilitates genetic counseling in these families. We would like to emphasize the need to perform DNA studies as a prerequisite for genetic counseling in affected families.


Glucose metabolism in the developing rat. Studies in vivo.
1. The specific radioactivity of plasma d-glucose and the incorporation of (14)C into plasma l-lactate, liver glycogen and skeletal-muscle glycogen was measured as a function of time after the intraperitoneal injection of d-[6-(14)C]glucose and d-[6-(3)H]glucose into newborn, 2-, 10- and 30-day-old rats. 2. The log of the specific radioactivity of both plasma d-[6-(14)C]- and d-[6-(3)H]-glucose of the 2-, 10- and 30-day-old rats decreased linearly with time for at least 60min after injection of labelled glucose. The specific radioactivity of both plasma d-[6-(14)C]- and d-[6-(3)H]-glucose of the newborn rat remained constant for at least 75min after injection. 3. The glucose turnover rate of the 30-day-old rat was significantly greater than (approximately twice) that of the 2- and 10-day-old rats. The relative size of both the glucose pool and the glucose space decreased with age. Less than 10% of the glucose utilized in the 2-, 10- and 30-day-old rats was recycled via the Cori cycle. 4. The results are discussed in relationship to the availability of dietary glucose and other factors that may influence glucose metabolism in the developing rat.


Cellular and humoral cross-immunity against two H3N2v influenza strains in presumably unexposed healthy and HIV-infected subjects.
Human cases of infection due to a novel swine-origin variant of influenza A virus subtype H3N2 (H3N2v) have recently been identified in the United States. Pre-existing humoral and cellular immunity has been recognized as one of the key factors in limiting the infection burden of an emerging influenza virus strain, contributing to restrict its circulation and to mitigate clinical presentation. Aim of this study was to assess humoral and cell-mediated cross immune responses to H3N2v in immuno-competent (healthy donors, n = 45) and immuno-compromised hosts (HIV-infected subjects, n = 46) never exposed to H3N2v influenza strain. Humoral response against i) H3N2v (A/H3N2/Ind/08/11), ii) animal vaccine H3N2 strain (A/H3N2/Min/11/10), and iii) pandemic H1N1 virus (A/H1N1/Cal/07/09) was analysed by hemagglutination inhibition assay; cell-mediated response against the same influenza strains was analysed by ELISpot assay. A large proportion of healthy and HIV subjects displayed cross-reacting humoral and cellular immune responses against two H3N2v strains, suggesting the presence of B- and T-cell clones able to recognize epitopes from emerging viral strains in both groups. Specifically, humoral response was lower in HIV subjects than in HD, and a specific age-related pattern of antibody response against different influenza strains was observed both in HD and in HIV. Cellular immune response was similar between HD and HIV groups and no relationship with age was reported. Finally, no correlation between humoral and cellular immune response was observed. Overall, a high prevalence of HD and HIV patients showing cross reactive immunity against two H3N2v strains was observed, with a slightly lower proportion in HIV persons. Other studies focused on HIV subjects at different stages of diseases are needed in order to define how cross immunity can be affected by advanced immunosuppression.


Influenza preparedness and response: involvement of African Field Epidemiology and Laboratory Training Programs, 2009.
The capacity of public health professionals to rapidly detect and respond to disease pandemics is critical to understand and control global disease spread. On June 11, 2009, the World Health Organization (WHO) declared H1N1 virus infection as pandemic. In May 2009, we assessed the participation of Field Epidemiology and Laboratory Training Programs (FELTPs) based in sub-Saharan Africa on pandemic influenza preparedness and response. We administered an electronic survey to directors and resident advisors of African Field Epidemiology Network (AFENET) member and associate FELTPs. The survey included questions on the following attributes: program involvement in suspected H1N1 investigations, experience in influenza outbreak investigations, national influenza surveillance and response plans, and H1N1 outbreak preparedness. Nine countries (100%) responded to the survey; all had existing national influenza response plans. Six programs reported their trainees had participated in past pandemic preparedness and response exercise, five (83%) of them were influenza specific. FELTPs played an important role in H1N1 surveillance and response in sub-Saharan Africa. Continued technical assistance and support to these programs is vital to foster their capacity to monitor and control public health threats.


Glucuronomannan GM2 from Saccharina japonica Enhanced Mitochondrial Function and Autophagy in a Parkinson's Model.
Parkinson's disease (PD), one of the most common neurodegenerative disorders, is caused by dopamine depletion in the striatum and dopaminergic neuron degeneration in the substantia nigra. In our previous study, we hydrolyzed the fucoidan from Saccharina japonica, obtaining three glucuronomannan oligosaccharides (GMn; GM1, GM2, and GM3) and found that GMn ameliorated behavioral deficits in Parkinsonism mice and downregulated the apoptotic signaling pathway, especially with GM2 showing a more effective role in neuroprotection. However, the neuroprotective mechanism is unclear. Therefore, in this study, we aimed to assess the neuroprotective effects of GM2 in vivo and in vitro. We applied GM2 in 1-methyl-4-phenylpyridinium (MPP+)-treated PC12 cells, and the results showed that GM2 markedly improved the cell viability and mitochondrial membrane potential, inhibited MPP+-induced apoptosis, and enhanced autophagy. Furthermore, GM2 contributed to reducing the loss of dopaminergic neurons in 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced mice through enhancing autophagy. These data indicate that a possible protection of mitochondria and upregulation of autophagy might underlie the observed neuroprotective effects, suggesting that GM2 has potential as a promising multifunctional lead disease-modifying therapy for PD. These findings might pave the way for additional treatment strategies utilizing carbohydrate drugs in PD.


Advanced nursing practitioners in primary care settings: an exploration of the developing roles.
Recent health care policies have resulted in patients having access to an integrated system of care that is quick and reliable. In concert with these changes, opportunities for professional development in nursing have increased, together with a reduction in the numbers of doctors. Advanced Nurse Practitioners (ANPs) have, therefore, developed to meet the complex demands of health care systems. This paper reports on a study that aimed to explore the current role of ANPs in primary care in the West Midlands region of the UK and how ANPs within three different nursing disciplines in primary care developed their roles over time. The study utilized a qualitative exploratory design incorporating a longitudinal element. Twenty-one ANPs were interviewed during phase one, 15 of whom were interviewed again during phase two, approximately 15 months later. Their managers (where appropriate) were also interviewed during phase one. The findings reveal that the nature and focus of practice varies between disciplines. At the extreme practice end of the practice-strategic continuum, Practice Nurse ANPs' expertise lies in their advanced practical assessment and diagnosis of individual patients, with little opportunity for strategic development. Health Visitor and District Nurse ANPs operate at the strategic end of the practice-strategic continuum, but operate differently at this level. Health Visitors, being community and public health focused are involved in multi-agency work, practice development and policy formulation. District Nurses work with individual patients/carers and the nursing team, thus their involvement in strategic developments tends to focus at the patient care level, such as protocol and practice developments, although their work also involves work in all three other domains. Overall, the findings reveal a unique role for all three with a potential career pathway for ANPs to become Nurse Consultants in the future.


Regio- and stereospecific synthesis of beta-sulfonamidodisulfides and beta-sulfonamidosulfides from aziridines using tetrathiomolybdate as a sulfur transfer reagent.
A comprehensive study of a general and effective one-step procedure for the synthesis of beta-sulfonamidodisulfides directly from N-tosyl aziridines in a regio- and stereospecific manner under neutral conditions without the use of any Lewis acid or base has been reported. This methodology is extended to the synthesis of an optically pure cyclic seven-membered disulfide 29. Synthesis of a variety of beta-sulfonamidosulfides involving tandem, multistep reactions in one pot is also reported.


A novel leader peptide which allows efficient secretion of a fragment of human interleukin 1 beta in Saccharomyces cerevisiae.
Killer strains of Kluyveromyces lactis secrete a toxin which presumably is processed during secretion from a larger precursor. Analysis of the sequence of the K. lactis killer toxin gene predicts that the first 16 amino acids at the amino terminus of the protein should represent its leader peptide. We have tested the capability of this leader peptide to direct secretion of a protein fused to it by inserting a synthetic oligonucleotide identical to the sequence of the putative leader peptide into a yeast expression vector. Subsequently, the cDNA coding for the secreted active portion of the human interleukin 1 beta (IL-1 beta) was fused to the leader peptide sequence of the killer toxin. This construction in Saccharomyces cerevisiae is capable of directing synthesis and secretion of correctly processed IL-1 beta into the culture medium.


An intracellular study of chemosensory fibers and endings.
1. The carotid body and its nerve, removed from anesthetized cats, were placed in physiological saline flowing under paraffin oil. The nerve, lifted into the oil, was used for either electrical stimulation or recording of the total afferent discharge. Intracellular recordings were obtained from individual nerve fibers and endings within the carotid body. The recording sites were identified by injecting Procion yellow through the intracellular electrodes; the tissues were then prepared for histology and observed with episcopic fluorescence or Nomarski optics. 2. Intracellularly recorded chemosensory fibers conducted at 1.1-30 m/s and usually displayed action potentials of regular amplitude. At times, however, some spikes become partially blocked while others maintained their original amplitude. "Natural" (hypoxia) or chemical (ACh or NaCN) stimulation induced different patterns of frequency changes of the large and small action potentials. This indicated nerve fiber branching at some distance from the recording site. 3. Intra- and extracellularly recorded spikes were blocked in 0 [Na+]0 by tetrodotoxin (TTX) or procaine. 4. During chemical stimulation, a slowly occurring depolarization (receptor or generator potential) was recorded intracellularly from the afferent fibers. It developed concomitantly with the increase in discharge. 5. Impalement of single nerve terminals (histologically identified) showed numerous "spontaneous" depolarizing potentials (SDPs) that had a mean amplitude of 5.6 mV, a mean duration of 46.1 ms, and nearly random distribution. They increased in frequency and summated during chemical stimulation. SDPs originated from either the site of recording or from neighboring areas. When the SDPs attained a certain amplitude, they seemed to give rise to action potentials. Also, relatively well developed or partially blocked spikes (apparently originating elsewhere) were recorded from single nerve terminals. 6. The receptor (generator) potential of chemosensory receptors appears to be an integrated response formed by multiple activity originating in different nerve endings.


Perspectives on Cannabis-Based Therapy of Multiple Sclerosis: A Mini-Review.
The consistency, efficacy, and safety of cannabis-based medicines have been demonstrated in humans, leading to the approval of the first cannabis-based therapy to alleviate spasticity and pain associated with multiple sclerosis (MS). Indeed, the evidence supporting the therapeutic potential of cannabinoids for the management of pathological events related to this disease is ever increasing. Different mechanisms of action have been proposed for cannabis-based treatments in mouse models of demyelination, such as Experimental Autoimmune Encephalomyelitis (EAE) and Theiler's Murine Encephalomyelitis Virus-Induced Demyelinating Disease (TMEV-IDD). Cells in the immune and nervous system express the machinery to synthesize and degrade endocannabinoids, as well as their CB1 and CB2 receptors, each mediating different intracellular pathways upon activation. Hence, the effects of cannabinoids on cells of the immune system, on the blood-brain barrier (BBB), microglia, astrocytes, oligodendrocytes and neurons, potentially open the way for a plethora of therapeutic actions on different targets that could aid the management of MS. As such, cannabinoids could have an important impact on the outcome of MS in terms of the resolution of inflammation or the potentiation of endogenous repair in the central nervous system (CNS), as witnessed in the EAE, TMEV-IDD and toxic demyelination models, and through other in vitro approaches. In this mini review article, we summarize what is currently known about the peripheral and central effects of cannabinoids in relation to the neuroinflammation coupled to MS. We pay special attention to their effects on remyelination and axon preservation within the CNS, considering the major questions raised in the field and future research directions.


Calmodulin-dependent protein kinases purified from rat brain and rabbit liver.
A calmodulin-dependent protein kinase was purified from rat brain by the same protocol used previously for a rabbit liver calmodulin-dependent glycogen synthase kinase. The rat brain kinase readily phosphorylated rabbit skeletal muscle glycogen synthase at sites 1b and 2, the same sites phosphorylated by rabbit liver calmodulin-dependent kinase. The two kinases have other similarities: substrate specificity, potent inhibition by sodium fluoride, and nearly equal Ka's (10-20 nM) for calmodulin. Also, both enzymes have similar Stokes radii, 70 A (rabbit liver) and 75 A (rat brain), but quite different sedimentation coefficients, 10.6 S and 17.4 S, respectively. Consequently, the calculated molecular weights are also different: 560,000 for the brain enzyme and 300,000 for the liver enzyme. The major subunit of the rat brain kinase appears to be a single 51-kDa peptide, not a doublet pattern of 51- and 53-kDa subunits that is characteristic of the rabbit liver enzyme. Our findings are consistent with the hypothesis that the rat brain and rabbit liver enzymes belong to a class of closely related calmodulin-dependent protein kinases, possibly isozymes. This class of enzymes may be responsible for regulating several of the known calcium-dependent physiological functions.


Administration of Apple Polyphenol Supplements for Skin Conditions in Healthy Women: A Randomized, Double-Blind, Placebo-Controlled Clinical Trial.
This clinical study was performed to evaluate the effects of continuous apple polyphenol (AP) administration on facial skin conditions and pigmentation induced by ultraviolet (UV) irradiation in healthy women participants. Participants (n = 65, age 20-39 years) were randomized to receive tablets containing AP (300 or 600 mg/day) or placebo in a double-blinded, placebo-controlled clinical trial. Continuous administration of AP for 12 weeks significantly prevented UV irradiation induced skin pigmentation (erythema value, melanin value, L value), although a dose-dependent relationship was not clearly observed. In contrast, no significant differences were detected between the groups with regard to water content and trans-epidermal water loss. Our study demonstrated that APs and their major active compounds, procyanidins, have several health benefits. Here, we report that continuous administration of AP for 12 weeks alleviated UV irradiation induced skin pigmentation, when compared with placebo, in healthy women.


A systematic survey of the sodium contents of processed foods.
Processed foods are major contributors to population dietary salt intake. Parts of the Australian food industry have started to decrease salt in a number of products. A definitive baseline assessment of current sodium concentrations in foods is key to targeting reformulation strategies and monitoring progress. Our objectives were to systematically collate data on the sodium content of Australian processed food products and compare sodium values against maximum target levels established by the UK Food Standards Agency (UK FSA). Categories of processed foods that contribute the majority of salt to Australian diets were identified. Food-composition data were sought for all products in these categories, and the sodium content in mg/100 g (or mg/100 mL for liquids) was recorded for each. Mean sodium values were calculated for each grouping and compared with the UK FSA benchmarks. Sodium data were collected for 7221 products in 10 food groups, 33 food categories, and 90 food subcategories. The food groups that were highest in sodium were sauces and spreads (1283 mg/100 g) and processed meats (846 mg/100 g). Cereal and cereal products (206 mg/100 g) and fruit and vegetables (211 mg/100 g) were the lowest in sodium. Sixty-three percent of food categories had mean sodium concentrations above the UK FSA targets, and most had wide ranges between the most and least salty product. Many products, particularly breads, processed meats, and sauces, have salt amounts above reasonable benchmarks. The variation in salt concentrations between comparable products suggests that reformulation is highly feasible for many foods.